We will use state-of-the-art, quantitative mass spectrometry-based proteomics to define membership of protein complexes in wild-type and mutant Mtb strains under a variety of growth and perturbational conditions. Systematic and unbiased definition of the membership of key protein complexes, and the changes in that membership under a variety of growth and perturbational conditions, will provide critical insights into the biological function of genes important for growth and survival of Mtb. By integrating results of proteomics, metablomics and transcriptomic analyses, a comprehensive picture of the interactions and expression programs critical for growth and survival of the organism should emerge. We will also use state-of-the-art, quantitative global proteomics to profile auxotrophic strains of Mtb to understand the roles of genes encoding proteins and non-coding RNAs of unknown function in M. tuberculosis. We will choose conditions/time points just before changes in growth occur to minimize the possibility that changes in protein abundance are a consequence of changes in growth and survival but, instead, are directly attributable to the mutation. Both wild type and mutant bacteria strains will be analyzed by comparative quantitative proteomic analysis. We anticipate that we will find several different types of changes. We will correlate changes with observations from other experiments across the consortium.

Public Health Relevance

Protein complexes, rather than individual proteins or binary interacting proteins, are the functional units in cells. The function of any protein is context dependent, deciphering the macromolecular context in which proteins are found is key to understanding cellular function and dynamics. The systematic unbiased ID of peptide/proteins under a range of conditions will provide critical insight into the pathways and survival of Mtb

National Institute of Health (NIH)
National Institute of Allergy and Infectious Diseases (NIAID)
Research Program--Cooperative Agreements (U19)
Project #
Application #
Study Section
Special Emphasis Panel (ZAI1)
Project Start
Project End
Budget Start
Budget End
Support Year
Fiscal Year
Total Cost
Indirect Cost
Harvard University
United States
Zip Code
Rego, E Hesper; Audette, Rebecca E; Rubin, Eric J (2017) Deletion of a mycobacterial divisome factor collapses single-cell phenotypic heterogeneity. Nature 546:153-157
Rock, Jeremy M; Hopkins, Forrest F; Chavez, Alejandro et al. (2017) Programmable transcriptional repression in mycobacteria using an orthogonal CRISPR interference platform. Nat Microbiol 2:16274
DeJesus, Michael A; Gerrick, Elias R; Xu, Weizhen et al. (2017) Comprehensive Essentiality Analysis of the Mycobacterium tuberculosis Genome via Saturating Transposon Mutagenesis. MBio 8:
Jansen, Robert S; Rhee, Kyu Y (2017) Emerging Approaches to Tuberculosis Drug Development: At Home in the Metabolome. Trends Pharmacol Sci 38:393-405
Kurthkoti, Krishna; Amin, Hamel; Marakalala, Mohlopheni J et al. (2017) The Capacity of Mycobacterium tuberculosis To Survive Iron Starvation Might Enable It To Persist in Iron-Deprived Microenvironments of Human Granulomas. MBio 8:
Mishra, Bibhuti B; Lovewell, Rustin R; Olive, Andrew J et al. (2017) Nitric oxide prevents a pathogen-permissive granulocytic inflammation during tuberculosis. Nat Microbiol 2:17072
Guinn, Kristine M; Rubin, Eric J (2017) Tuberculosis: Just the FAQs. MBio 8:
Köster, Stefan; Upadhyay, Sandeep; Chandra, Pallavi et al. (2017) Mycobacterium tuberculosis is protected from NADPH oxidase and LC3-associated phagocytosis by the LCP protein CpsA. Proc Natl Acad Sci U S A 114:E8711-E8720
Botella, Helene; Vaubourgeix, Julien; Lee, Myung Hee et al. (2017) Mycobacterium tuberculosis protease MarP activates a peptidoglycan hydrolase during acid stress. EMBO J 36:536-548
Baric, Ralph S; Crosson, Sean; Damania, Blossom et al. (2016) Next-Generation High-Throughput Functional Annotation of Microbial Genomes. MBio 7:

Showing the most recent 10 out of 28 publications